Scientific Research Journal (SCIRJ), Volume II, Issue XII, December 2014

ISSN 2201-2796

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Phenotypical Morphometry Variation of Aedes

Aegyti in Manado
Wulan PJ Kaunang
Student in Doctoral of Entomology, Post-Graduate Program
The University of Sam Ratulangi
Manado

Jantje Pelealu
Study Program of Entomology, Post-Graduate Program
The University of Sam Ratulangi
Manado

Max Tulung
Study Program of Entomology, Post-Graduate Program
The University of Sam Ratulangi
Manado

Christin Salaki
Study Program of Entomology, Post-Graduate Program
The University of Sam Ratulangi
Manado

Abstract- Aedes aegypti (Linne, 1762) is the main vector of

arboviruses, namely yellow fever, dengue and Chikungnya. In
North Sulawesi it has found variation in morphology, ecology,
behavior and vector competence. Vector control is still the best
way to prevent dengue fever. Understanding the morphological
characteristics that become characters in the phenotypic
mosquito is very important in the efforts of controlling and
classification. This study aims to find morphological variation of
Aedes aegypti in the city of Manado. This research applies
descriptive research techniques with direct experiments in the
laboratory. The morphologic character of mosquito samples
from selected locations were then measured by using a
stereomicroscope
KH8700
completed
with
cameras,
measurement software of morphological characters and
statistical programs. The morphometry data were then analyzed
with multivariate and factor analysis by using IBM SPSS 20. The
results showed that of the twelve morphological characters were
analyzed, it was found that Aedes spp from 4 areas in the city of
Manado has undergone phenotypical characters variations
(morphology), characterized by the formation of five clusters or
groups based on the degree of similarity in morphometry. There
are three groupings of morphological characters which are
variance factor of 1: (5.941 / 12) x 100% = 49.508%, variance
factor of 2: 1.798 / 12) x 100% = 14.98%, and variance 3 factors:
(1,184 / 12 x 100% = 9.86%.
Index Terms variation, morphometry, Aedes aegypti,
Manado

I. INTRODUCTION
Aedes is a mosquito kind normally found in the tropics. Its
name is derived from the Greek word Aedes, which means "not
nice", because this mosquito spreads dangerous diseases, such
as dengue fever and yellow fever. Aedes albopictus sp. is a
common species in Asia. His legs are black and white stripes.

Aides sp. also known as the spreads of dengue and yellow

fever. Aedes aegypti sp. and Aedes albopictus sp. are found in
the tropics and sub-tropics. In Asia it is on the first rank in
terms of the number of patients with Dengue Fever in each
year. Meanwhile, from 1968 to 2009, the World Health
Organization (WHO) noted that Indonesia as the country with
the highest number of cases on Dengue Fever in Southeast
Asia. Of the total number of such cases, about 95% cases
occured in children below 15 years.
Aedes aegypti breeds in water reservoirs, such as bathtub,
jars, drums, flower vases, and used items that can be container
of rain water in urban and suburban areas. Aedes albopictus,
likewise, but they are usually more widely available outside of
houses. Aedes aegypti mosquitoes prefer to bite on sheltered
areas, such as around the house. Biting activity reaches the top
during the light intensity change. They can bite all day and the
highest is before sunset. Short flight range is 50-100 meters,
except in the wind. Mosquitoes will rest in dark and cool places
after they suck blood, until the blood absorption for egg
development completed. Mosquitoes will find a runny place to
put and lay their eggs, and then the mosquitoes will begin to
look for blood again for the next spawning cycle.
From January to August this year it occurred 324 cases,
with 11 of them ended the children death. The area with the
highest number of cases, namely Districts of Malalayang,
Wanea, and Tikala (Depkes Manado, tribunsnews.com). In
East Bolaang Mongondow, along the current year local health
department has recorded 23 cases, two of whom died. They
were residents of the Bai village and citizens of Kotabunan.
The highest number of cases occurred between May and July,
in Nuangan District as the highest number of dengue cases
territory.

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Scientific Research Journal (SCIRJ), Volume II, Issue XII, December 2014
ISSN 2201-2796

Aedes spp is the main vector of dengue virus result in

dengue fever. Several studies have found a bionomic change of
dengue mosquitoes in some areas. Changes in mosquito
behavior can affect the mosquitoes morphology, The changes
are caused by the habitat shift, climate change, and some
efforts that eradicate mosquitoes with insecticide that
stimulates mosquito resistance reactions.
Morphological characteristics or phenotype is the
expression of genes that are affected by environment.
Individual species pphenotype resulted from genotype (genes)
and environment influence these organisms. Substantial
phenotypic variation in a population is due to differences in
genotype. Variations derived from DNA mutation, migration
between populations (gene flow), and changes in the
composition of genes through recombination in sexual
reproduction. Although there are variations that occur
continuously through these processes, most of the species
genome is identical in all individuals of that species. (1)
Genetic variation is the variation due to genetic changes
(mainly mutations) and passed on to offspring through the cell
nucleus in the gametes. (2) Environmental variation is the
variation caused by environmental changes, while the genetic
material remains and is not inherited. Small changes in
genotype can lead to dramatic changes in the phenotype.
Changes in gene frequency in a population can occur rapidly
when there is human intervention. Several factors that can
change gene frequencies in a population are mutation,
migration between populations, genetic drift and selection.
The influences of these four factors determine the genes
frequency and population characteristics, and then randomly
passed on to the next generation.
In Africa it has been found variation in the morphology,
ecology, and behavior and vector competence. Even Aedes
aegypti has been divided into Aedes aegypti (Aaa) that is
specific domestic (living) live in life settlements area /
anthropophilic and cosmotropical distribution with lighter /
brighter body color/morphology, while Aedes aegypti
formosus (Aaf) which is endemic to Africa is darker and its
habitat is farther from the domestic (Paupy et. al. 2010).
Rondonuwu reported that there has been a shift in habitat
between Aedes albopitus that inhabit marginal areas near the
forest area towards domestic / urban in the Manado city.
One common method used to determine the phenotypical
variation of species populations is a morphometric analysis.
Morphometry can be defined as the method described
morphologic characters through measurement, calculation or
scoring (Bookstein and Strauss, 1982). Morphometry can be
applied to determine the particular species kinship, the various
species differentiation, and variations in the species and to
identify a species. Some morphometric studies have been done,
such as morphometry of Apis dorsata Binghami and Apis
nigrocincta Smith (Mokosuli, 2013), morphometric variation of
Papilio polythes L (Makhzuni et.al, 2013), and morphometric
analysis of Anopheles subpictus (Chillar, 2014).
Based on the literature study conducted by the authors,
there is only a few published research reports on morphological

16

variation of Aedes spp. Morphological variation study needs to

be done, because with the increase in dengue cases, the efforts
to eradicate mosquitoes as vectors intensified. Generally the
mosquito eradication uses insecticide. Insects are animals that
have a high degree of genetic adaptation to various conditions
or have a high adaptive capability. Routine insecticide
treatments will bring Aedes variants resistant to insecticides,
such as Anopheless spp which has high resistance levels to
organoclorin and organophosphates insecticides in Indonesia
today. Resistance is formed due to the mosquitos
physiological reactions to adapt or survive. Resistance is also
influenced by genetic factors or genetic modification as a
mosquitoes adaptation form. Genetic modification will have
implications on mosquitoes phenotypes or morphology. Until
now there has been no research study report of the
morphological variation of Aedes albopictus in Manado. This
study aims to find out morphological variation of Aedes spp in
Manado City. This study will then be equipped with barcode
genetic studies using mitochondrial DNA CO1 gene.
II. MATERIALS AND METHODS
Research Location and Time
Mosquito samples obtained from several locations in
Manado the city, namely Malalayang, Winangun, Kairagi, and
Tuminting. The sample areas can be clustered based on the
height of the sea surface. Each region was taken 50 adult
mosquitoes without classifying males or females.
Tools and Materials
Tools used were pyrex glassware, three-dimensional stereo
microscope of hirox KH-8700 models and stereo microscope
of Carl Zeizz "STEMI DV 4" stereomicroscope. Both
microscopes are equipped with CPUs, monitors, cameras, and
measurement software of bee organs. The material used
consists of 70% alcohol, chloroform p.a., cotton, and distilled
water. Adult mosquito samples were obtained in domestic area.
Research procedures
a. Sampling Mosquitoes
Alive mosquitoes terminated with 70% alcohol. Mosquito
samples from each location is stored in the sample bottle with
70% alcohol and placed in the sample box to maintain the
temperature of 200-250C. Mosquitoes further sorted and only
mosquitoes that have complete body parts (not defective /
broken) during an arrest or preparation used for morphological
analysis.
b. Morphometry analysis
The mosquitoes morphological analysis used 3-dimensional
robotic stereo Mikroscope of KH700 hirox completed with
morphological measurement software, camera, and statistical
programs. Morphological characters were measured according
to the Andrew and Bar method (2013); Rueda, (2008), namely
the head, antenna, proboscis, Torax, wings, legs and abdomen
structures (Figure 1).

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Scientific Research Journal (SCIRJ), Volume II, Issue XII, December 2014
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Fig. 1. Morphology Parameters used for morphometric analysis (Rueda LM. 2008)

c. Data Analysis Techniques

Measuring the length and width of the morphological
characters used the software on the microscope. Measurement
result data of morphological characters analyzed by Cluster and
factorial used IBM SPSS 20.
III. RESULTS AND DISCUSSION
Grouping Morphometry Characters
To determine the grouping of morfometerical characters
used multivariate analysis technique that is cluster analysis
with IBM SPSS 20. Cluster analysis application traditionally
aimed to explore and establish a classification / taxonomy
empirically. Since its ability to partition, the cluster analysis

can be applied widely. Cluster analysis aims to classify objects

or characters, so that each object or character that has the
closest similarity to the other is in the same cluster. Clusters
that are formed have a high internal homogeneity and high
external heterogeneity. Cluster analysis is used to process the
insect morphometry data with classifying species aim based on
similar morphometry or classifying certain morphometric
characters based on morphometry similarity (Mokosuli, 2013;
Seenivasagan et. Al., 2009; Chhilar, 2014). A total of 11
clusters that were formed on the cluster analysis of
morphometric characters of Aedes spp obtained in 4 areas in
Manado city on 12 Aedes morphological characters measured
(Figure 2).

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Fig. 2. Morphological Character Cluster of Aedes spp in Manado

Clusters that are formed subsequently manifested in a

dendogram to determine the grouping of properties /
morphological characters based on the similarity degree. In
dendogram analysis results obtained morphological characters
of the head length (PK), the antenna length (Pant), the
abdomen width (Labd), tarsus length (PTR), the tarsus width
(LTR), the front wing width (LSD) and a long proboscis (ppb)
of 4 samples regions to form one cluster (the first cluster).
Therefore, it has very close similarity level of mosquitoes
morphology from 4 regions on these characters. LTR character

has similarities with the character of the rear leg femur length
(PFkb) with ecludian distance 5, thus forming the cluster itself
(second cluster). LTR and PFkb character has similarities with
of the abdomen length (Pabd) character with ecludian distance
18, thus forming a third cluster. Character of the front wing
length (PSD) has a very close resemblance to the character of
the rear leg tarsus length (Ptkb), thus forming a fourth cluster.
The last formed cluster is LTR characters, PFkb, and Pabd with
farthest ecludian distance 25 (Figure 3).

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Fig. 3. Grouping Dendogram of Morphology Character on Aedes obtained from 4 regions in Manado city.

Relationships among Morphology Characters

To obtain the relationship form of morphometric variables
used factor analysis. Factor analysis is a statistical analysis
technique that aims to explain the relationship structure
between the variables observed by raising a number of factors
or components or latent variables less than a number of
variable origins. The main purpose of factor analysis is to
explain the relationships structure among many variables in the
form factor or latent or formations variables.
Factor analysis with PCA method used IBM SPSS 20.
There are three factors (components) with the eigenvalues
values of 5.941; 1.798; and 1,184 (Appendix 1). Eigenvalues
values indicate the variable numbers that become factor
members. Accordingly, PCA analysis morphometry of Aedes
bee ... on factor of one consisting of 5.941 variables, factor of
two consisting of 1.798 variables, and factor of three consisting

of 1,184 variables. Of the 12 components (variables) were

included in the factors analysis that each variable has variance
1, the total variance is 12 x 1 = 12. In accordance with the
number of factors that are formed and the amount of variance
of each variable that is known, it can further explained that
each factor as well as overall form factor: Variance factor of 1:
(5.941 / 12) x 100% = 49.508%, Variance factor of 2: 1.798 /
12) x 100% = 14.98%, and Variance factor of 3: (1,184 / 12 x
100% = 9.86%. This means that 49.50% of the factor variance
variability of one can be explained by this factor, and
respectively 14.98% and 9.86% can be explained by a factor of
two and a factor of three. Accordingly, these three factors can
explain 73.95% of the variability of 12 morphological
characters of Aedes spp.

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Component Matrixa
Component
1
2
PK
-.096
.373
Pant
.619
.616
PTR
.673
.441
LTR
.824
.006
PSD
.883
-.202
LSD
.564
-.535
Pabd
.958
-.116
Labd
.296
-.764
Ppb
.864
-.246
PFkb
.833
.150
Ptkb
.321
.124
Ptrkb
.878
.249
Extraction Method: Principal
Analysis.
a. 3 components extracted.

20

3
.732
-.031
-.091
-.016
.026
.056
-.097
.303
.000
.143
.671
-.252
Component

Image Grouping variables from three factors formed.

Fig. 4. Aedes sp sample from Manado

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Fig. 5. Samples of Aedes albopictus from Airmadidi

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Fig. 7. Characters Morphometry of Aedes Morphology

Fig. 8. Front legs and abdomen of Aedes sp from Kairagi

Fig. 6. Comparison of the Aedes Head from Malalayang and Kairagi

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Scientific Research Journal (SCIRJ), Volume II, Issue XII, December 2014
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Morphological variation of Aedes sp in an area widely

studied io different areas of endemic dengue. Morphological
variation in tarsomer character was found in Aedes simpsoni in
Uganda, Kenya, and North Africa. Midtarsomer 1 and 2 have a
longer band or strip than the male and female adult mosquito
on anthropophilic and non-anthropophilic population in Kenya
and Uganda. Besides, on tarsomer also be found in scutal
character variation, middle leg claws (Lutwama and Mukwaya,
1994). Paupy et. al. (2010) found variations in the pale scale
character of Aedes aegypti abdominal in Niakhar, Senegal.
This variation depends on the season and nesting places of
mosquito larvae. Aedes has the ability to perform a striking
morphological variation, although it is still located in the same
living area (biotypes). Aedes in Manado has shown some
variations in morphological characters. Variations in
morphology (phenotypical) should be followed up in the
genetic analysis using specific DNA gene as a barcode or
standard to see the genetic variation rate in the mosquitoes that
live in the same biotype. Morphological modifications in the
adaptation response to pressure changes of the environment,
climate, habitat, and defense against insecticides is a common
response to the species, in the class of insects that are known to
have high adaptability. Morphological variations of Aedes spp
become an important study, because Aedes ecological role as a
dengue virus vector causes hemorrhagic fever and viral
chikugunya. Morphological modifications based on the
evolution theory have two possibilities, namely progressive or
retrogressive. In this study, morphological modifications
actually lead to the progressive evolution or the species
member formation in a more stable population. In another
study conducted by the author, the dengue cases prevalence in
Manado city has increased, therefore morphological
modifications do no direct mosquito population member to the
inability to survive or adapt (retrogressive).
IV. CONCLUSION
Of the twelve morphological characters analyzed, it has
been found that Aedes spp from 4 areas in Manado city has
undergone phenotypical character variation (morphology). It is
characterized by five clusters or groups formation based on the
morphometry similarity degree. There are three groupings of
morphological characters, namely factor variation of 1: (5.941 /
12) x 100% = 49.508%, factor variation of 2: 1.798 / 12) x

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100% = 14.98%, factor variation of 3: (1,184 / 12 x 100% =

9.86%.
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